New Developments in High Performance Magnetic Separation Technology for Laboratory and Industrial Applications

Transcription

1 New Developments in High Performance Magnetic Separation Technology for Laboratory and Industrial Applications David Humphries, Martin Pollard D.O.E. Joint Genome Institute/ Lawrence Berkeley National Laboratory

2 Emerging Magnetic Technology for Genomic, Proteomic and Bio-Medical Applications Hybrid 384-well magnet plates for molecular separation Hybrid 96-well magnet plates New development areas from deep well plates to single molecule manipulation Technology transfer and industry collaboration

3 Unique Technology with Broad Applicability New class of magnet plates developed at JGI/LBNL for high-throughput purification of biological samples in highdensity microtiter plates Technology can selectively separate proteins, DNA and other molecules from various contaminates based solely on a magnetic field Magnetic structure is a unique hybrid of permanent magnet and ferromagnetic materials that produces magnetic fields significantly stronger than those of commercially available magnet plates

4 Hybrid Magnet with Plate Interface and 384-Well Microtiter Plate

5 High Throughput and Benchtop Use Hybrid magnetic structures are currently an enabling technology for sequencing approximately 2.5 Gigabases of DNA per month at the DOE Joint Genome Institute. Hybrid magnet plates have been developed for general bench top use in addition to automated high throughput applications.

6 Computer Model of Hybrid MagneticStructure Ferromagnetic Poles Permanent Magnet Material

7 High Fields and Strong Gradients 600 Force Density and Gradient Plot 500 T^2/m and T/m Y (mm) Grad(T/m) Force Density (T^2/m)

8 Target Species Are Magnetized Magnet plates work in conjunction with 0.5 to 5.0 micron magnetic beads Beads are attached to DNA or other targets such as proteins by means of specialized bead coatings Various bead types are commercially available

9 Applications Functional genomics Genetic sequencing Proteomics Immunological drug screening Automated DNA purification Automated protein purification Any magnetic bead based purification method Single molecule manipulation

10 BiomekFX Liquid Handling Robot with Four Hybrid Magnets Installed

11 Close-up of Hybrids on Biomek-FX Deck

12 Advantages of Hybrid Technology Fields are 70 to 100% stronger than that of the best commercial magnet plates measured Produces higher sample yields and faster processing times Greater holding power means more robust process Compatible with most 96, 384 and well standard microtiter plates

13 General Attributes These hybrid structures are energized by permanent magnets and require no external power source They are compact, with a footprint slightly larger than a standard microtiter plate and a thickness of approximately 1 to 1.8 inches Documented fabrication techniques have been developed that allow for high quantity/quality production of these structures

14 Performance Comparison Relative field strengths of five different magnet plates were compared. Three of the magnet plates were developed at JGI/LBNL. The other two magnet plates are commercially available models. The field strengths were measured at two heights: (a) at the magnet surface and (b) at 1 cm above the magnet surface. Measurements were made using a Hall effect probe.

15 Fields at magnet surface are 70% greater than industry 384-well magnet plates tested Field Strength at Magnet - Industry vs JGI/LBNL Field at Magnet (G.) Commercial 96-Well Commercial 384-Well Perforated Bar-LBNL Bar Magnet -LBNL Hybrid-LBNL Magnet Type

16 Fields at magnet surface are 70 to 100% greater than industry 384-well magnet plates tested Field Strength at Magnet Pole Tips - Industry vs JGI/LBNL Field at Magnet (G.) Commercial 96-Well Commercial 384-Well LBNL Perf. Bar LBNL Bar Magnet Magnet Type LBNL Hybrid LBNL 96-Well Hybrid

17 Fields 1 cm above the magnet are approx. 400% stronger than 384-well commercial magnet Field Strength 1 cm Above Magnet Industry vs JGI/LBNL Field 1 cm Up (G.) Commercial 96- Well Commercial 384-Well Perforated Bar- LBNL Bar Magnet- LBNL Hybrid-LBNL Magnet Type

18 Drawdown Performance Comparison Video still sequence provides a draw-down comparison between the hybrid and a common commercial magnet plate 1 micron beads are suspended in a viscous solution in 384-well microtiter plates Non-magnetic mock-up plate is included for visual reference

19 Hybrid vs Commercial Plate - Initial Conditions

20 Drawdown After 20 Seconds

21 Drawdown After 30 Seconds

22 Drawdown After 50 Seconds

23 96-Well, 384-Well & 1536-Well Industry-Standard Microtiter Plates

24 New Hybrid-96 Prototype Plates In Testing Second generation Hybrid structures Optimized for higher fields and gradients Initial configurations for 96 well microtiter plates Designed for fast drawdown and beneficial distribution of targets

25 Hybrid 96-well Prototype

26 Computer Model of 96-Well Hybrid Fields 1.1 Tesla Fields At Pole Tips

27 Second Generation 96-well Hybrid Magnetic Structure w/ 96-well Microplate

28 Second Generation 96-well Hybrid Magnetic Structure w/o Microplate

29 Hybrid-96 Core Magnetic Structure

30 Hybrid-96 Exploded View of Core Magnetic Structure

31 Hybrid-96 Detail View of Pole Tips

32 Microtiter Plate Well to Pole Relationship

33 Continuing and Future Work Continued development of the hybrid concept for higher performance and broader applicability Fabrication of new, long period device for 1 and 2 ml deep well plates Hybrid variants for single molecule manipulation under rapid development Bio-medical applications of Hybrid technology in conceptual stages

34 Long Period Hybrid w/ 2 ml Deep Well Plate

35 Long Period Hybrid w/ Upper Interface

36 Long Period Hybrid w/o Upper Interface

37 Long Period Hybrid Exploded View

38 Technology Transfer Opportunities Hybrid magnetic technology being patented by the University of California and Lawrence Berkeley National Laboratory Hybrid technology available for transfer to industry through the Tech Transfer Department of LBNL LBNL evolving as a center for advanced magnetic R&D for genomic, proteomic and bio-medical applications LBNL open to collaborative development efforts as part of its tech transfer process

39 Advanced Fabrication Facilities at Lawrence Berkeley National Laboratory

40 State-of-the-Art High Precision Machine Tools

41 This work was performed under the auspices of the US Department of Energy's Office of Science, Biological and Environmental Research Program by the University of California, Lawrence Livermore National Laboratory under Contract No. W ENG-48, Lawrence Berkeley National Laboratory under contract No. DE-AC03-76SF00098 and Los Alamos National Laboratory under contract No. W ENG-3. LBNL-57845